- 期刊
- OpenAccess
利用PCR選殖菊花rDNA之內轉錄間隔區
Cloning of Internal Transcribed Spacer Region of rDNA in Dendranthema grandiflorum by Polymerase Chain Reaction
摘要
設計一組引子（primer），序列分別為IT1-5' CGTAACAAGGTTTCC3'和IT2-5' AGTTTCTTCTCCTCC3'，可有效複製6個菊花品種（系）的5.8S核糖體核酸（rRNA）基因與內轉錄間隔區（internal transcribed spacer，ITS），經聚合脢連鎖反應（polymerase chain reaction，PCR）複製產物長度為747 bp。將其中一品種的內轉錄間隔區進行定序，扣除兩端引子長度（30 bp），總長為717 bp，與其他高等植物的相同區域比較，發現PCR產物有27 bp屬於18SrRNA基因的3'端，與52 bp屬於26S rRNA基因的5'端。因此，內轉錄間隔區實長為638 bp，此區包含ITS1、5.8S rRNA基因與ITS2等三區，其長度分別為253 bp、164 bp及221 bp。由上述研究証實，本研究之引子可藉由PCR技術，將各菊花品種（系）rDNA之ITS有效選殖，其中ITS1及ITS2為變異快速之DNA區域，可以由此獲得許多分子標誌，供未來應用於品種鑑別及新品種保護上。
並列摘要
Using polymerase chain reaction (PCR), the entire nucleotide sequences of internal transcribed spacer (ITS) region between 18S and 26S ribosomal RNA (rRNA) genes were amplified among six cultivars/lines of Dendranthema grandiflorum Ramat. Two primers for PCR, IT1-5' CGTAACAAGGTTTCC 3' and ITS2-5' AGTTTCTTCTCCTCC 3' were designed for amplification and sequencing. The length of PCR products including primers (30 bp) was 747 bp. To compare with other higher plant species, this sequence contained 27 bp of 18S rRNA gene, ITS region, and 52 bp of 26S rRNA gene. Therefore, the length of ITS region of D. grandiflorum is 638 bp in which 253 bp of ITS1, 164 bp of 5.8S rRNA gene, and 221 bp of ITS2 are present.